| Budget Amount *help |
¥2,300,000 (Direct Cost: ¥2,300,000)
Fiscal Year 1995: ¥1,200,000 (Direct Cost: ¥1,200,000)
Fiscal Year 1994: ¥1,100,000 (Direct Cost: ¥1,100,000)
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| Research Abstract |
In order to analyze interactions between the bacterial flagellar switch proteins and other proteins, it is necessary to purify the switch proteins in native form. In particularly, the FliM protein formed inclusion bodies when it is overproduced. Even after purification of the protein after denaturing with guanidine hydrochloride or urea, it formed inclusion bodies. To purify FliM in native form, we tried to remove urea from the sample with dialysis just after denaturing and succeeded to purify. Also, we found that this protein gradually form oligomers in neutral pH.On the other hand, we studied properities of interaction between FliM and the chemotaxis signaling protein CheY in detail. In the case of FliN,we can purify it in large amount and high purity. Therefore, we tried to isolate crystals of FliN and found conditions for formation of pseudocrystals. We will continue to work to find conditions for FliN crystals for X-ray crystallography. In studies on interaction of the switch proteins with the MS ring, we use new techniques, one is quick-freeze deep-etch replica images observed by electron microscopy and the other is atomic force microscopy. By the former we found a new structure in the cytoplasmic surface of the MS ring consisting of the switch proteins. By the latter, we examined force profiles of the MS ring complex with or without the FliG protein and found that there were drastic differences.
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